PXD018520

PXD018520 is an original dataset announced via ProteomeXchange.

Title	Carboxypeptidase B-assisted charge-based fractional diagonal chromatography for deep screening of C-terminome
Description	The determination of protein C-termini is of great significance for protein function annotation and proteolysis re-search. However, the progress of C-terminomics is still far behind its counterpart, N-terminomics, because of the low reactivity of the carboxyl group. Herein, we developed a negative selection strategy, termed carboxypeptidase B-assisted charge-based fractional diagonal chromatography (CPB-ChaFRADIC), to achieve a global C-terminome analysis. The highly reactive carboxypeptidase B cleavage was utilized to reduce the charge state of non-C-terminal peptides. Together with high-performance charge-based frac-tional diagonal chromatography, the C-terminal peptides could be isolated. Such a strategy was also applied for profiling C-termini from Escherichia coli cell lysates, and 441 canonical C-termini and 510 neo-C-termini originating from proteolytic processing were identified. These findings represent 2-fold and 5.8-fold that of identified C-termini via direct analysis, respectively. Using parallel digestion with trypsin and LysC, such a strategy enabled the identification of 604 canonical C-termini and 818 neo-C-termini, rep-resenting the largest C-terminome dataset of E. coli, and no deficiency in His/Lys/Arg-containing C-terminal peptides was observed. The presented CPB-ChaFRADIC strategy is therefore a highly efficient and unbiased strategy for large-scale C-terminome analysis. Furthermore, using the CPB-ChaFRADIC strategy, we identified 87 cleavage sites and 82 substrates of caspase-3 in Jurkat cells, demonstrating that the CPB-ChaFRADIC strategy shows great promise in promoting proteolysis research.
HostingRepository	PRIDE
AnnounceDate	2020-05-25
AnnouncementXML	Submission_2020-05-24_22:43:05.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Lingfan Chen
SpeciesList	scientific name: Escherichia coli; NCBI TaxID: 562; scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	S-carboxamidoethyl-L-cysteine; 6x(13)C labeled residue; monohydroxylated residue
Instrument	Q Exactive

Revision	Datetime	Status	ChangeLog Entry
0	2020-04-14 02:19:15	ID requested
⏵ 1	2020-05-24 22:43:06	announced

Chen L, Shan Y, Yang C, Sui Z, Zhang X, Zhang L, Zhang Y, Carboxypeptidase B-Assisted Charge-Based Fractional Diagonal Chromatography for Deep Screening of C-Terminome. Anal Chem, 92(12):8005-8009(2020) [pubmed]

submitter keyword: c-terminomic,ChaFRADIC,caspase,cleavage site

Lihua Zhang
contact affiliation	Dalian Institute of Chemical Physics, Chinese Academy of Sciences
contact email	lihuazhang@dicp.ac.cn
lab head
Lingfan Chen
contact affiliation	Dalian Institute of Chemical Physics
contact email	anxuanmuma@126.com
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD018520
     1. Label: PRIDE project
     2. Name: Carboxypeptidase B-assisted charge-based fractional diagonal chromatography for deep screening of C-terminome